Supercritical fluid-assisted controllable fabrication of open and highly interconnected porous scaffolds for bone tissue engineering
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Recently tremendous progress has been evidenced by the advancements in developing innovative three-dimensional (3D) scaffolds using various techniques for addressing the autogenous grafting of bone. In this work, we demonstrated the fabrication of porous polycaprolactone (PCL) scaffolds for osteogenic differentiation based on supercritical fluid-assisted hybrid processes of phase inversion and foaming. This eco-friendly process resulted in the highly porous biomimetic scaffolds with open and interconnected architectures. Initially, a 23 factorial experiment was designed for investigating the relative significance of various processing parameters and achieving better control over the porosity as well as the compressive mechanical properties of the scaffold. Then, single factor experiment was carried out to understand the effects of various processing parameters on the morphology of scaffolds. On the other hand, we encapsulated a growth factor, i.e., bone morphogenic protein-2 (BMP-2), as a model protein in these porous scaffolds for evaluating their osteogenic differentiation. In vitro investigations of growth factor loaded PCL scaffolds using bone marrow stromal cells (BMSCs) have shown that these growth factor-encumbered scaffolds were capable of differentiating the cells over the control experiments. Furthermore, the osteogenic differentiation was confirmed by measuring the cell proliferation, and alkaline phosphatase (ALP) activity, which were significantly higher demonstrating the active bone growth. Together, these results have suggested that the fabrication of growth factor-loaded porous scaffolds prepared by the eco-friendly hybrid processing efficiently promoted the osteogenic differentiation and may have a significant potential in bone tissue engineering.
Keywordssupercritical foaming polycaprolactone bone tissue engineering osteogenic differentiation bone morphogenic protein-2
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This work was supported by the National Natural Science Foundation of China (U1605225, 31570974, and 31470927), the Public Science and Technology Research Funds Projects of Ocean (201505029), the Promotion Program for Young and Middle-aged Teacher in Science and Technology Research of Huaqiao University (ZQN-PY107) and the Program for Innovative Research Team in Science and Technology in Fujian Province University.
Compliance and ethics The author(s) declare that they have no conflict of interest.
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- An, J., Teoh, J.E.M., Suntornnond, R., and Chua, C.K. (2015). Design and 3D printing of scaffolds and tissues. Engineering 1, 261–268.Google Scholar
- Cai, Y., Tong, S., Zhang, R., Zhu, T., and Wang, X. (2018). In vitro evaluation of a bone morphogenetic protein-2 nanometer hydroxyapatite collagen scaffold for bone regeneration. Mol Med Report 17, 5830.Google Scholar
- Chen, B., Kankala, R.K., Chen, A., Yang, D., Cheng, X., Jiang, N., Zhu, K., and Wang, S. (2017). Investigation of silk fibroin nanoparticledecorated poly(L-lactic acid) composite scaffolds for osteoblast growth and differentiation. Int J Nanomed 12, 1877–1890.Google Scholar
- Chen, C., Liu, Q., Xin, X., Guan, Y., and Yao, S. (2016). Pore formation of poly(ε-caprolactone) scaffolds with melting point reduction in supercritical CO2 foaming. J Supercrit Fluids 117, 279–288.Google Scholar
- Choudhury, M., Mohanty, S., and Nayak, S. (2015). Effect of different solvents in solvent casting of porous pla scaffolds—in biomedical and tissue engineering applications. J Biomater Tissue Eng 5, 1–9.Google Scholar
- Davies, O.R., Lewis, A.L., Whitaker, M.J., Tai, H., Shakesheff, K.M., and Howdle, S.M. (2008). Applications of supercritical CO2 in the fabrication of polymer systems for drug delivery and tissue engineering. Adv Drug Deliver Rev 60, 373–387.Google Scholar
- Delmote, J., Teruel-Biosca, L., Gómez Ribelles, J.L., and Gallego Ferrer, G. (2017). Emulsion based microencapsulation of proteins in poly(Llactic acid) films and membranes for the controlled release of drugs. Polym Degrad Stabil 146, 24–33.Google Scholar
- Deng, A., Chen, A., Wang, S., Li, Y., Liu, Y., Cheng, X., Zhao, Z., and Lin, D. (2013). Porous nanostructured poly-L-lactide scaffolds prepared by phase inversion using supercritical CO2 as a nonsolvent in the presence of ammonium bicarbonate particles. J Supercrit Fluids 77, 110–116.Google Scholar
- Diaz-Gomez, L., Concheiro, A., Alvarez-Lorenzo, C., and García-González, C.A. (2016a). Growth factors delivery from hybrid PCLstarch scaffolds processed using supercritical fluid technology. Carbohyd Polym 142, 282–292.Google Scholar
- Diaz-Gomez, L., Yang, F., Jansen, J.A., Concheiro, A., Alvarez-Lorenzo, C., and García-González, C.A. (2016b). Low viscosity-PLGA scaffolds by compressed CO2 foaming for growth factor delivery. RSC Adv 6, 70510–70519.Google Scholar
- Duarte, A.R.C., Mano, J.F., and Reis, R.L. (2009). Perspectives on: supercritical fluid technology for 3D tissue engineering scaffold applications. J Bioact Compat Polym 24, 385–400.Google Scholar
- Fanovich, M.A., Ivanovic, J., Misic, D., Alvarez, M.V., Jaeger, P., Zizovic, I., and Eggers, R. (2013). Development of polycaprolactone scaffold with antibacterial activity by an integrated supercritical extraction and impregnation process. J Supercrit Fluids 78, 42–53.Google Scholar
- Hile, D.D., Amirpour, M.L., Akgerman, A., and Pishko, M.V. (2000). Active growth factor delivery from poly(D,L-lactide-co-glycolide) foams prepared in supercritical CO2. J Control Releas 66, 177–185.Google Scholar
- Jing, X., Mi, H., Cordie, T., Salick, M., Peng, X., and Turng, L.S. (2014). Fabrication of porous poly(ε-caprolactone) scaffolds containing chitosan nanofibers by combining extrusion foaming, leaching, and freeze-drying methods. Ind Eng Chem Res 53, 17909–17918.Google Scholar
- Kankala, R.K., Zhang, Y., Wang, S., Lee, C.H., and Chen, A. (2017b). Supercritical fluid technology: an emphasis on drug delivery and related biomedical applications. Adv Healthc Mater 6, 1700433.Google Scholar
- Kankala, R.K., Zhu, K., Sun, X., Liu, C., Wang, S., and Chen, A. (2018a). Cardiac tissue engineering on the nanoscale. ACS Biomater Sci Eng 4, 800–818.Google Scholar
- Kankala, R.K., Chen, B., Liu, C., Tang, H., Wang, S., and Chen, A. (2018c). Solution-enhanced dispersion by supercritical fluids: an ecofriendly nanonization approach for processing biomaterials and pharmaceutical compounds. Int J Nanomed 13, 4227–4245.Google Scholar
- Krause, B., Mettinkhof, R., van der Vegt, N.F.A., and Wessling, M. (2001). Microcellular foaming of amorphous high-T g polymers using carbon dioxide. Macromolecules 34, 874–884.Google Scholar
- Lee, S.J., Lee, D., Yoon, T.R., Kim, H.K., Jo, H.H., Park, J.S., Lee, J.H., Kim, W.D., Kwon, I.K., and Park, S.A. (2016). Surface modification of 3D-printed porous scaffolds via mussel-inspired polydopamine and effective immobilization of rhBMP-2 to promote osteogenic differentiation for bone tissue engineering. Acta Biomater 40, 182–191.PubMedGoogle Scholar
- Lian, Z., Epstein, S.A., Blenk, C.W., and Shine, A.D. (2006). Carbon dioxide-induced melting point depression of biodegradable semicrystalline polymers. J Supercrit Fluids 39, 107–117.Google Scholar
- Mao, J., Duan, S., Song, A., Cai, Q., Deng, X., and Yang, X. (2012). Macroporous and nanofibrous poly(lactide-co-glycolide)(50/50) scaffolds via phase separation combined with particle-leaching. Mater Sci Eng-C 32, 1407–1414.Google Scholar
- Mathieu, L.M., Montjovent, M.O., Bourban, P.E., Pioletti, D.P., and Månson, J.A.E. (2005). Bioresorbable composites prepared by supercritical fluid foaming. J Biomed Mater Res 75A, 89–97.Google Scholar
- Moshiri, A., and Oryan, A. (2012). Role of tissue engineering in tendon reconstructive surgery and regenerative medicine: current concepts, approaches and concerns. Hard Tissue 1, 11.Google Scholar
- Nam, Y.S., Yoon, J.J., and Park, T.G. (2015). A novel fabrication method of macroporous biodegradable polymer scaffolds using gas foaming salt as a porogen additive. J Biomed Mater Res 53, 1–7.Google Scholar
- Qu, X., Cao, Y., Chen, C., Die, X., and Kang, Q. (2014). A poly(lactide-coglycolide) film loaded with abundant bone morphogenetic protein-2: a substrate-promoting osteoblast attachment, proliferation, and differentiation in bone tissue engineering. J Biomed Mater Res 103, 2786–2796.Google Scholar
- Rajabzadeh, S., Liang, C., Ohmukai, Y., Maruyama, T., and Matsuyama, H. (2012). Effect of additives on the morphology and properties of poly (vinylidene fluoride) blend hollow fiber membrane prepared by the thermally induced phase separation method. J Membrane Sci 423–424, 189–194.Google Scholar
- Salerno, A., Clerici, U., and Domingo, C. (2014a). Solid-state foaming of biodegradable polyesters by means of supercritical CO2/ethyl lactate mixtures: towards designing advanced materials by means of sustainable processes. Eur Polymer J 51, 1–11.Google Scholar
- Salerno, A., Fanovich, M.A., and Pascual, C.D. (2014b). The effect of ethyl-lactate and ethyl-acetate plasticizers on PCL and PCL-HA composites foamed with supercritical CO2. J Supercrit Fluids 95, 394–406.Google Scholar
- Salerno, A., Diéguez, S., Diaz-Gomez, L., Gómez-Amoza, J.L., Magariños, B., Concheiro, A., Domingo, C., Alvarez-Lorenzo, C., and García-González, C.A. (2017). Synthetic scaffolds with full pore interconnectivity for bone regeneration prepared by supercritical foaming using advanced biofunctional plasticizers. Biofabrication 9, 035002.PubMedGoogle Scholar
- Tomasko, D.L., Li, H., Liu, D., Han, X., Wingert, M.J., Lee, L.J., and Koelling, K.W. (2003). A review of CO2 applications in the processing of polymers. Ind Eng Chem Res 42, 6431–6456.Google Scholar
- Woodruff, M.A., and Hutmacher, D.W. (2010). The return of a forgotten polymer—polycaprolactone in the 21st century. Prog Polymer Sci 35, 1217–1256.Google Scholar
- Yano, K., Hoshino, M., Ohta, Y., Manaka, T., Naka, Y., Imai, Y., Sebald, W., and Takaoka, K. (2009). Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by Escherichia coli and dimerized by biochemical processing. J Bone Miner Metab 27, 355–363.PubMedGoogle Scholar
- Zhao, G., Cao, Y., Zhu, X., Tang, X., Ding, L., Sun, H., Li, J., Li, X., Dai, C., Ru, T., et al. (2017). Transplantation of collagen scaffold with autologous bone marrow mononuclear cells promotes functional endometrium reconstruction via downregulating ΔNp63 expression in Asherman’s syndrome. Sci China Life Sci 60, 404–416.PubMedGoogle Scholar